In criminal technology, identifying bloodstain species is essential in determining the nature of a case and investigation direction. This study explores spectral nondestructive species identification method. Using CRI Nuance spectral imager to carry MISytem 3.0 spectral image analysis software, in the spectrum range of 450 nm?950 nm, 180 groups of objects including human blood trace samples, animal blood trace samples, and mixed human and animal blood trace samples on 30 carriers were analyzed by spectral image analysis. Results were analyzed using the slope difference between the samples’ two substances. The hyperspectral imaging revealed that the three types of bloodstains could be distinguished on most carriers, with discrimination rate exceeding 70%. This study supplements the bloodstains species identification method and serves as a reference for an optical nondestructive bloodstain examination.